br Experimental design materials and methods X

Experimental design, materials and methods X-ray diffraction datasets were collected at synchrotron beamlines ID23-1 [2] (ESRF) and XALOC [3] (ALBA), from flash cooled (in liquid nitrogen) single crystals and kept under nitrogen gas during data collection. Diffraction data already reduced and scaled, in the form of structure factors, were deposited in the Protein Data Bank under accession codes PDB: 4d5u and PDB: 4jfb and are freely retrievable to the community. Structure factors were submitted to the anisotropy server at UCLA for analysis (http://services.mbi.ucla.edu/anisoscale/) (Fig. 1B) [4]. The server outputs the diffraction intensity falloff as a function of sars-cov for the three main direction of the reciprocal space; vectors a in red, b* in green and c* in blue. The same structure factors were analyzed using the Phenix software package [5] to visualize diffraction data along the (l,k) and (l,h) planes to exemplify the diffraction anisotropy (Fig. 1C). h, k and l are the Miller indices of the reflections along the a*, b* and c* vectors, respectively. The intensity falloff is clearly seen as k and h increase, confirming a stronger diffraction along increasing l (c*). Fig. 1A links the real cell with the reciprocal space. The crystal packing is depicted with the asymmetric unit made of two OmpF trimers colored in blue. From the arrangement of the OmpF trimers, it is clear that most of the crystal contacts are along c, with the contacts along a and b being mediated by detergents, and thereby being weaker. The reciprocal space is shown with each reciprocal vector in the same color as the one used by the anisotropy server. The reciprocal cell values have been determined using the online server, http://www.ruppweb.org/new_comp/reciprocal_cell.htm, and their size have been multiplied by 5000 to be able to visualize them next to the real cell. For ease of visualization, the origin of the reciprocal cell has been offset to the right of the origin of the real cell. These data exemplify a stronger diffraction along c*, which is also the direction with the most crystal contacts, along c.
Acknowledgments This work was supported by the ANR grant EMMA-10-049-01, 13-BSV5-0001-01 and the “labellisation par la Ligue Nationale Contre le Cancer”. The authors wish to thank Mike Sawaya for useful discussions during the preparation of these data. We acknowledge the synchrotrons SOLEIL, ESRF and ALBA for allocation of beam time and we are thankful to the beamline staff for their support and for the extensive beam-time required for membrane-protein projects. Financial support of Arnaud Kilburg was received from the Rhône-Alpes region. The crystallization work benefited from the Protein Science Facility of the SFR Biosciences Lyon.
Value of the data
Data Sunflower pollen is an important source of inhalant allergens. In this data article we are sharing the mass spectrometry data for identification of seven allergens from sunflower pollen. This dataset is associated with the article “Search for allergens from the pollen proteome of sunflower (L.): A major sensitizer for respiratory allergy patients” [1].
Experimental design, materials and methods Using bottom-up proteomic approach we have previously identified sunflower pollen allergens [1]. The experimental workflow (illustrated in Fig. 1) was designed to identify allergenic pollen proteins of sunflower. First, clinico-immunological tests were performed to understand the prevalence of sensitivity towards sunflower pollen among the atopic population. Sera from selected sunflower positive patients were used as the probe to detect the IgE-reactive proteins from two-dimensional electrophoretic separated proteome of sunflower pollen. Finally, these allergens were identified by mass-spectrometry.
Acknowledgments Authors are thankful to Mr. Chanchal Chakraborty, Mr. Amarendranath Biswas, Smt. Kaveri Ghosh and Mr. Jadav Ghosh for their technical support. NG, GS and BS receive funding from Indian Council of Medical Research (Grant no. 3/1/3/WL/JRF-2011/HRD-102 (41416) Dated 19.3.2012), Council of Scientific and Industrial Research (Grant no. 03/015(0414)/2011-EMR-I) and University Grant Commission (Grant no. F.2-8/2002(SA-I)). We are thankful to all organizations that funded our research.